PUBLICATION
Photoinduced RNA interference using DMNPE-caged 2'-deoxy-2'-fluoro substituted nucleic acids in vitro and in vivo
- Authors
- Blidner, R.A., Svoboda, K.R., Hammer, R.P., and Monroe, W.T.
- ID
- ZDB-PUB-080421-8
- Date
- 2008
- Source
- Molecular Biosystems 4(5): 431-440 (Journal)
- Registered Authors
- Svoboda, Kurt
- Keywords
- none
- MeSH Terms
-
- Animals
- Cells, Cultured
- Deoxyribonucleotides/pharmacology*
- Embryo, Nonmammalian/cytology
- Embryo, Nonmammalian/drug effects
- Fluorine*
- Gene Silencing/radiation effects
- Light*
- Nitrobenzenes/pharmacology*
- RNA Interference*/radiation effects
- Zebrafish
- PubMed
- 18414741 Full text @ Mol. Biosyst.
Citation
Blidner, R.A., Svoboda, K.R., Hammer, R.P., and Monroe, W.T. (2008) Photoinduced RNA interference using DMNPE-caged 2'-deoxy-2'-fluoro substituted nucleic acids in vitro and in vivo. Molecular Biosystems. 4(5):431-440.
Abstract
Various chemical modifications to RNA have been incorporated in attempts to improve their pharmacological properties for RNAi interference (RNAi). Recent studies have shown that small interfering RNA (siRNA) containing 2'-fluoro modifications can elicit gene silencing through RNAi. Despite developments in using chemical modifications for increased stability, safety, and efficiency of these therapeutics, they still face challenges of spatial and temporal targeting. One potential targeting strategy is to use photocaging techniques, which involve the covalent attachment of photolabile compounds to the effector nucleic acid species that block bioactivity until exposed to near UV light. In this study we demonstrate that fully 2'-fluorinated nucleic acids (FNAs) can be caged for photoactivated gene silencing in cell culture and in zebrafish embryos. This strategy combines the improvement in chemical and enzymatic stability associated with 2'-substitutions with the targeting ability of a photoinducible trigger. Statistical alkylation of FNAs with 1-(4,5-dimethoxy-2-nitrophenyl)diazoethane (DMNPE) improved resistance to enzymatic degradation, reduced RNAi effectiveness, and protected the biological system from toxic doses of the effector. Photo-exposure to 365 nm light partially restored the silencing activity of the 2'-fluoro siRNAs. These results suggest that photocaging may offer control over RNAi therapeutics for spatially and temporally directed activation, while improving enzymatic stability and potentially enabling therapeutic dosing via light dose intensity.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping